Quantification of sofalcone in human plasma and urine by high performance liquid chromatography-mass spectrometry

Abstract

Sofalcone, isolated from the root of the Chinese medicinal plant Sophora subprostrata, is well known to be a mucosal protective agent for gastritis and peptic ulcer treatment. Although the LC-MS/MS and HPLC-DAD methods for assay of plasma concentration of sofalcone were reported before for the pharmacokinetic study, they were either too complicated or not sensitive enough for current pharmacokinetic study. In addition, no urinary assay method or pharmacokinetic information was available. Thus an improved high performance liquid chromatography-mass spectrometric method employing negative electrospray ionization was developed for the determination of sofalcone concentration in human plasma and urine sample. A liquid-liquid extraction method was utilized to extract sofalcone together with the indometacin (internal standards) from 0.5ml of human plasma or urine samples. Multiple reaction monitoring was used for quantification by monitoring the transition of m/z from 449.5 to 313.1 for sofalcone and 356.9 to 313.0 for IS. The validation of the method regarding to specificity, sensitivity, linearity, reproducibility, accuracy and stability was evaluated. The lower limit of quantification (LLOQ) of the developed assay method for sofalcone was 0.5ng/ml and the linear calibration curve was acquired with R(2)>0.99 between 0.5 and 500ng/ml for both plasma and urine samples. The intra- and inter-day variations of the current assay were evaluated with the relative standard deviation (RSD) within 13.77% at low concentration of quality control samples (QCs) and 8.71% for other QCs, whereas the mean accuracy ranged from 96.21 to 107.33%. The samples were found to be stable under the storage conditions at least for a month and other experimental conditions. This validated method was then utilized to test sofalcone concentration in clinical samples. Based on these data, the pharmacokinetic behavior of sofalcone in plasma as well as urine was described. As a conclusion, the present method proved to be a rapid and sensitive analytical tool for sofalcone in human plasma or urine samples and has been successfully applied to a clinical pharmacokinetic study of in healthy Chinese subjects.